Biogenic Amines and Microbiological Profile of Egyptian Cheeses
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Vol. 8(3), pp. 130-139, March 2014 DOI: 10.5897/AJFS08.059 ISSN 1996-0794 Copyright © 2014 African Journal of Food Science Author(s) retain the copyright of this article http://www.academicjournals.org/AJFS Full Length Research Paper Biogenic amines and microbiological profile of egyptian cheeses Khaled Meghawry El-Zahar Food Science Department, Faculty of Agriculture, Zagazig University, 44511 Zagazig, Egypt. Received 14 December, 2013; Accepted 17 February, 2014 Cheeses are among those high-protein-containing foodstuffs in which enzymatic and microbial activities cause the formation of biogenic amines (BAs) from amino acids decarboxylation. Most of the methods for amine determination in these products involve acid extraction followed by a liquid-liquid purification step to selectively separate amines and amino acids. This study aimed to describe the development of biogenic amines in Egyptian cheeses during ripening and storage regimes. Biogenic amines content in Mish, Ras and Blue cheeses were 270-1300, 340-980 and 210-700 mg/kg, respectively. The dominant biogenic amines were different. This work confirms that the main biological feature influencing amines formation is the extent of growth of microorganisms, like Enterococci, characterized by decarboxylase activity. It is important to note that the presence of biogenic amines due to the activities of these microorganisms is maintained within safe levels. In Egypt, reports dealing with the Egyptian cheeses (Mish, Ras and Blue) are scanty. So, the present work was carried out to fill the gap in our knowledge on its microbiological and biochemical features, focusing on hygiene and consumer health aspects. Key words: Biogenic amines, food safety, proteolysis, ripening, Enterococci spp. INTRODUCTION Milk and milk products are very important in human which they are responsible for many essential functions. nutrition and, among them; cheese is considered a good They can be naturally present in many foods such as source of proteins, vitamins and minerals. However, fruits and vegetables, meat, fish, chocolate and milk, but cheese is one of the most fermented foods commonly they can also be produced in high amounts by associated with biogenic amines (BAs) contamination. microorganisms through the activity of amino acid These compounds are basic nitrogenous compounds decarboxylases (Ten Brink et al., 1990). Excessive formed by series of microorganisms, mainly by consumption of these amines can be of health concern decarboxylation of amino acids or “in vivo” also by de- because when there is are no equilibrate assumption in amination and trans-amination of aldehydes and ketones human organism, can generate different degrees of (Loizzo et al., 2012, 2013). Biogenic amines are diseases determined by their action on nervous, gastric compounds commonly present in living organisms in and intestinal systems and blood pressure (Suzzi and *Corresponding author. E-mail: [email protected]. Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution License 4.0 International License El-Zahar 131 Gardini, 2003). Biogenic amines are low molecular weight (Wöhrl et al., 2004). nitrogenous bases, they were found in fermented foods The ability of microorganisms to decarboxylate amino and cheese (Mohamed et al., 2013). acid is highly variable. Due to strain-specific, it is impor- Also, biogenic amines are low-molecular nitrogenous tant to count decarboxylase-positive microorganisms to compounds that are formed in foodstuffs mainly by estimate; the risk of BAs food content and to prevent BAs microbial decarboxylation of the precursor amino acids accumulation in food products. Presence and accumula- (Alberto et al., 2002). The importance of observing BAs tion of BAs depends on many factors such as presence content lies in potential toxicity to human, mainly when of specific bacteria (Enterococci, Micrococci, the concentration is up to 100 mg/kg (or up to 100 mg/L). Enterobacteriaceae and Lactobacilli) and enzymes, Thus, the presence of BAs significantly influences the availability of free amino acids, presence of suitable food quality and safety (Smit et al., 2005). cofactors, that is, pH level, water activity, temperature The presence of relevant amounts of BAs in cheeses and salt content, type of cheese, ripening and storage has been documented (Martuscelli et al., 2005; Kung et period (Galgano et al., 2001). Some controversial results al., 2007; Pintado et al., 2008; Ladero et al., 2009; have been reported on the contribution of Enterococci sp. Mercogliano et al., 2010). In cheeses, BAs formation is in BAs production in cheeses, and in particular in hista- caused by curdling and cheese decarboxylase-positive mine (Sumner and Taylor, 1989). Enterococci have a microorganisms. Histamine (HIS), tyramine (TYR), long history of use as artisanal cultures for preparation of putrescine (PTR), cadaverine (CAD), spermidine (SPD), various types of cheeses (Izquierdo et al., 2009), they are spermine (SPR), tryptamine (T), and β-phenylethylamine sometimes associated with pathogenicity (Khan et al., (PE) are frequently found in these products. Cheese is 2010), can cause endocarditic, bacteraemia, and several one of the fermented foods most commonly associated infections, as well as multiple antibiotic resistances with BAs poisoning; mainly HIS, TYR, PTR and CAD. (Kayser, 2003). Although, the chemical composition and Indeed, the term ‘‘cheese reaction” to refer to it (Ten microbiological quality of cheeses in Egyptian markets Brink et al., 1990). Tyramine and histamine are the most have been studied extensively, little data is available on abundant and frequent BAs in cheese (Fernández et al., the occurrence of biogenic amines in Egyptian cheeses. 2007). Consumption of food containing high levels of BAs Therefore, this survey was undertaken to determine the is considered undesirable since it can be associated with presence of BAs in commercially available cheeses several toxicological problems such as respiratory during ripening and storage, also to make an assessment distress, headache, hyper- or hypo-tension or allergies of the health hazard arising from the consumption of (Ladero et al., 2010). These problems are especially these products especially by susceptible individuals. severe in consumers with low levels of the enzymes Other studies such as Lorencová et al. (2012) and involved in the detoxification system (mono and di-amine Buňková et al. (2010) deal with selection and study of oxidases), either by genetic disorders (Caston et al., microorganism (such as lactic acid bacteria), which are 2002) or medical treatments (Halász et al., 1994). The major producer of biogenic amine. However, these works content of biogenic amines and polyamines significantly explore the particular biogenic amine production in differed according to the technology of ripening. The growth medium, where the concentration of biogenic cheeses unwashed during ripening had much higher amine could be biased optimal environment for the contents of all observed amines and polyamines in bacteria metabolisms. Moreover, some strains of the comparison with the washed-rind cheeses. The mean starter lactic acid bacteria (such as Lactococcus lactis content of putrescine, cadaverine and tyramine exceeded subsp. lactis) have decarboxylase activity that was 100 mg/kg in unwashed-rind cheeses, while the other observed in model environment of growth broth. amines occurred at lower levels. The content of all Behaviour of these strains has not been investigated in detected amines was very low in washed-rind cheeses; real system of the cheese and can be different in no tryptamine, phenylethylamine and histamine were comparison with condition in growth broth. found. The effect of storage on the amine formation was The objective of our pilot study was to compare the not confirmed (Samková et al., 2013). BAs content and other selected parameters in Egyptian Physiologically, histamine is one of the most effective cheese and to review hypothesis that the BAs content BAs; it has vasoactive and psychoactive effects (Repka- developed during the ripening and storage period is Ramirez and Baraniuk, 2002). Moreover, it is the main related to the presence of decarboxylase positive strain BAs involved in food poisoning and it is limited in some of Enterococci sp. foodstuffs by law. At non-toxic doses, food borne histamine can cause intolerance symptoms such as MATERIALS AND METHODS diarrhoea, hypotension, headache, pruritus and flushes. Just 75 mg of histamine, a quantity commonly present in Cheese samples some meals, can induce symptoms in the majority of healthy persons with no history of histamine intolerance A total 85, 49 and 44 of Mish, Ras and Blue cheeses samples were 132 Afr. J. Food Sci. purchased from different Egyptian retail markets and small scale Microbiological analysis factories. The samples collected were 6-48 months old. They were kept in sterile plastic bags and transported to the laboratory of Food For each cheese sample, 10 g was weighed and dispersed asep- Science Department, Zagazig University (Egypt), then stored at 4 tically in 90 mL of citrate buffer (2%,w/v) and homogenized in a ±1°C until analyzed. sterile polyethylene bag using a Stomacher (Seward Laboratory Blender Stomacher 400 Lab Blender UK) for 1.5 min. Serial dilu- tions were made in 0.1% sterile peptone water and all determina- Chemical analysis tions were made in triplicates (Messer et al., 1985). The enumera- tion